Eucalyptus globules belonging to the Myrtaceae family was explored for the synthesis of zinc oxide nanoparticles and for biological applications. The aqueous extract of the synthesized zinc nanoparticles (ZnNPs) was characterized using UV-visible spectrophotometer, FTIR, SEM and TEM. The aqueous broth was observed to be an efficient reducing agent, leading to the rapid formation of ZnNPs of varied shapes with sizes ranging between 52–70 nm. In addition, antifungal activity of the biosynthesized ZnNPs was evaluated against major phytopathogens of apple orchards. At 100 ppm of ZnNPs, the fungal growth inhibition rate was found to be 76.7% for Alternaria mali, followed by 65.4 and 55.2% inhibition rate for Botryosphaeria dothidea and Diplodia seriata, respectively. The microscopic observations of the treated fungal plates revealed that ZnNPs damages the topography of the fungal hyphal layers leading to a reduced contraction of hyphae. This considerable fungicidal property of ZnNPs against phytopathogenic fungi can have a tremendous impact on exploitation of ZnNPs for fungal pest management and ensure protection in fruit crops.